Upload
others
View
3
Download
0
Embed Size (px)
Citation preview
Pediatrics Hematology and OncologyLab of SCT and Immunotherapy Johann Wolfgang Goethe-University60596 Frankfurt / Main
Umesoft GmbHSteinbacher Str. 14-24
65760 Eschborn
Multivariate Analysis of the immunologic
system´s reconstitution after an Allogeneic
Stem Cell Transplantation
Emilia Salzmann & Melanie Bremm
Umetrics User Meeting - 2011
Allogeneic Stem Cell Transplantation (SCT)
219.08.2011
Search: HLA-matched related or unrelated donor
Graft:- Bone Marrow- PBSC- Cord Blood
Patient Conditioning
Antonio Pezzutto, Taschenatlas der Immunologie, Thieme Verlag, 2006
Transplantation
Umetrics User Meeting - 2011
Immune Reconstitution post-SCT
319.08.2011
Chemotherapy
SCT
-10 0 +10 +20 +30 +40
Cells / µl
5000
0
Cell-Regeneration
New immune cells are
built from donor stem
cells
Immuno-therapy
Days
Immuno-therapy
Relapse
Umetrics User Meeting - 2011
Immune Reconstitution post-SCT
419.08.2011
• A rapid immune reconstitution following SCT is of
central importance to protect patients from relapse
and infectious complications
• An adequate immune reconstitution can efficiently
eliminate recipient’s residual malignant cells
• Patients with delayed immune reconstitution show
an enhanced probability of relapse and severe
infections
Umetrics User Meeting - 2011
Therapeutic Intervention
519.08.2011
• Intervention is mainly based on
minimal residual disease (MRD)*
diagnostic and donor/recipient
chimerism
Unfortunately part of the patients
already exhibit a manifest relapse at
this time point
Today’s standard
*Evidence for the presence of residual malignant cells
• Identify patients following SCT with
an increased risk of relapse and/or life-
threatening infections on the basis of
immune reconstitution
Introducing prophylactic cell-
therapies (for ex. DLI, NK cell
donation) may be beneficial for high-
risk patients to overcome their
susceptibility for relapse or infectious
complications
Future aim
Umetrics User Meeting - 2011
Study Design
619.08.2011
Aim:
Generate a multivariate model to divide leukemia patients following SCT
into risk-groups
Strategy:
1) Choose adequate leukocyte subpopulations to use for model
generation
2) Design a multivariate model on the basis of age-matched leukocyte
subpopulation values of 100 healthy children and adolescents
3) Classify the immune reconstitution of the patients post allo-SCT on
the basis of the new model
4) Create risk-groups on the basis of the classification result
Umetrics User Meeting - 2011
Dimensionality Reduction
719.08.2011
p[1]
p[2]
Leukocytes
Lymphocytes
CD3+ (T cells)
CD3+CD4+ (helper T cells)
CD3+CD8+ (cytotoxic T cells)
CD3-CD56+ (NK cells)
CD19+ (B cells)
CD14+ (Monocytes)
Input Result
Dimensionality
Reduction (PCA)
p[3]
Umetrics User Meeting - 2011
Reference Model
819.08.2011
Age-matched
lymphocyte
subpopulation
reference values
from 100 healthy
children are used to
generate a
multivariate model.
This model defines
the range of normal
lymphocyte values.
t[1]t[2]
t[3]
3-component multivariate model:
Umetrics User Meeting - 2011
Prediction
919.08.2011
Classification: Do immune-status values of transplanted
children reach the reference ellipsoid?
Evaluation of leukocyte subsets of 32 pediatric
patients undergoing allogeneic SCT
Umetrics User Meeting - 2011
Prediction - Example
1019.08.2011
Monitoring Time (Days Following SCT)
tPS[1]
tPS[2]
tPS[3]a
c
T2
Ra
ng
e V
alu
esb
tPS[1]
tPS[3]
tPS[2]
M. Koenig et al., BMT, 2009
Umetrics User Meeting - 2011
Overall Survival
1119.08.2011
a b
Classification at day 200 Classification at day 300
Significantly higher number of long-time survivors among
the low-risk group compared to the high-risk group
Umetrics User Meeting - 2011
Hierarchical model
1219.08.2011
PCA
Leukocytes
Lymphocytes
CD3+ (T cells)
CD3+CD4+ (helper T cells)
CD3+CD8+ (cytotoxic T cells)
CD3-CD56+ (NK cells)
CD19+ (B cells)
CD14+ (Monocytes)
to 3 – component
Model(t1, t2, t3)
OPLST2-Range Risk-Index
Distance from the centre of the ellipsoid
Umetrics User Meeting - 2011
Risk- Index
1319.08.2011
the risk Index to classify a given patient's life expectancy
"an event" or "a non-event" is:
- non-event patients : - risk-index = 0 : For long-time survivors the event-risk converges to zero
- event-patients : - risk-index > 1 : patient who dies within the first year
- risk-index = 1 : patient who dies exactly in 1 year
- risk-index < 1 : patient who dies after 1 year
Umetrics User Meeting - 2011
Early prediction
1419.08.2011
• 2 component OPLS-model
(orthogonal PLS)
Result• Data from first 3 months to predict 1
year
Training data-Set: 16 patients with
acute leukemia
Use time dependence variables T2 -
dd for modeling risk-index . Where dd
is the day post SCT
First scenario
0,0
0,1
0,2
0,3
0,4
0,5
0,6
0,7
0,8
0,9
Com
p[1
]P
Comp No.
Model (OPLS/O2PLS) R2Y(cum)
Q2(cum)
SIMCA-P+ 12.0.1 - 2011-06-01 11:14:42 (UTC+1)
Umetrics User Meeting - 2011
Prediction 1 year from the first 3 months
1519.08.2011
Umetrics User Meeting - 2011
Prediction 1 year from the first 3 months
1619.08.2011
Umetrics User Meeting - 2011
Prediction
1719.08.2011
Training-Set (n= 16) predict-event predict-no-event
Event in 1st. year 3 3
No Event 1 9
Prediction-Set (n= 46) predict-event predict-no-event
Event in 1st. year 7 7
No Event 9 19
Data from first 3 months to predict 1 year
Umetrics User Meeting - 2011
Loading PLot
1819.08.2011
0,00
0,10
0,20
0,30
0,40
0,50
0,60
0,70
-0,02 0,00 0,02 0,04 0,06 0,08 0,10 0,12 0,14 0,16 0,18 0,20 0,22 0,24 0,26 0,28 0,30 0,32 0,34 0,36 0,38 0,40 0,42
po[1
]
pq[1]
Model (OPLS/O2PLS)pq[Comp. 1]/po[Last comp.]
R2X[1] = 0,499719 R2X[XSide Comp. 1] = 0,395335
14
2733
4150
61
6977
86
96
SIMCA-P+ 12.0.1 - 2011-06-07 17:55:37 (UTC+1)
Umetrics User Meeting - 2011
Thank You for Your Attention
1919.08.2011
PD Dr. U. Köhl
Dr. R. Esser
Dr. S. Klöß
Dr. S. Hünecke
C. Brehm
E. Auth
A. Quaiser
S. Wehner
R. Müller
S. Erben
R. El Kalaäoui
T. Gardlowski
S. Betz
O. Zimmermann
Lab
of
SC
T a
nd
Im
mu
no
thera
py
Jo
han
n W
olf
gan
g G
oeth
e-U
niv
ers
ity Prof. E. Falkenberg
Prof. M. Behl
Prof. A. Orth
E. Salzmann
Un
ivers
ity o
f A
pp
lied
Scie
nces F
ran
kfu
rt
Prof. A. Orth
E. Salzmann
Um
eso
ft G
mb
H
This project was supported by ‘DFG (GRK-1172)’, ‘BMBF (FKZ 01FP09120B)’,
‘Frankfurter Stiftung für krebskranke Kinder’ and ‘Hilfe für krebskranke Kinder Frankfurt e.V.’
Umetrics User Meeting - 201120
19.08.2011
Umetrics User Meeting - 2011
Flowcytometric Monitoring
2119.08.2011
Umetrics User Meeting - 2011
CD3 PC5
CD
8 E
CD
Lymph + Check FSC Lymph
Flowcytometric Measurement
2219.08.2011
CD45 FITC
SS
Gate: ungated
CD3 PC5
Lymph + Check FSC Lymph
SS
FS
Gate: Lymph
CD
14
PC
7
CD45+
SS CD3 PC5
CD
56
PE
Lymph + Check FSC Lymph
CD3 PC5
CD
19
EC
D
Lymph + Check FSC Lymph
CD
4 P
E
CD3 PC5
Lymph + Check FSC Lymph
Umetrics User Meeting - 2011
Age Correction
2319.08.2011
11 years
11 years
T-H
elp
er/µ
l
Age (months)
Age (months) Sqrt(Age (months))
log(
T-H
elp
er/µ
l+1
)
log(
T-H
elp
er/µ
l+1
)
Umetrics User Meeting - 2011
Infections/ GvHD post-SCT
2419.08.2011
GvHD Grade IV
Umetrics User Meeting - 2011
Example
2519.08.2011
Pat-ID
PS.ObsID.Obs.
ID..Dauer..
M1.T2Range
PS.1...3. Risk_Index...
L-3 10 52,8594 1,26736
L-3 13 76,7786 1,26736
L-3 21 23,8722 1,26736
L-3 28 40,7368 1,26736
L-3 35 27,2886 1,26736
L-3 41 74,0817 1,26736
L-3 49 31,5839 1,26736...
L-7 15 463,007 0,404208
L-7 19 312,14 0,404208
L-7 27 178,683 0,404208
L-7 36 110,954 0,404208
L-7 48 93,8538 0,404208
L-7 54 149,43 0,404208
L-7 61 102,21 0,404208
L-7 75 104,068 0,404208
L-7 83 105,946 0,404208
Obs ID
(Primary)
Obs ID
(Number)
M1.YPred[1]
(Risk_Index
_M1_14)
M1.YVar(Risk
_index_M1_1
4)
L-2 1 0,903135 1,07353
L-3 2 0,521172 1,26736
L-5 3 0,230717 0
L-6 4 0,50773 0
L-7 5 1,02757 0,404208
L-11 6 0,566901 0,34795
L-13 7 0,780602 0
L-14 8 1,61624 1,81592
L-15 9 0,599637 0
L-16 10 0,639127 0
L-18 11 0,320025 1,34686
L-19 12 0,331459 0
L-21 13 1,54062 1,77184
L-23 14 1,35444 2,26708
L-108 15 0,391192 0,732932
L-110 16 -0,482898 0
Primary ID Number
M1.T2Rang
ePS.1...3._
M1_14
M1.T2Rang
ePS.1...3._
M1_27
M1.T2Rang
ePS.1...3._
M1_33
M1.T2Rang
ePS.1...3._
M1_41
M1.T2Rang
ePS.1...3._
M1_50
M1.T2Rang
ePS.1...3._
M1_61
M1.T2Rang
ePS.1...3._
M1_69
M1.T2Rang
ePS.1...3._
M1_77
M1.T2Rang
ePS.1...3._
M1_86
M1.T2Rang
ePS.1...3._
M1_96
Phase M1 M1 M1 M1 M1 M1 M1 M1 M1 M1
Maturity 14 27 33 41 50 61 69 77 86 96
Source
M1.T2Rang
ePS.1...3.
M1.T2Rang
ePS.1...3.
M1.T2Rang
ePS.1...3.
M1.T2Rang
ePS.1...3.
M1.T2Rang
ePS.1...3.
M1.T2Rang
ePS.1...3.
M1.T2Rang
ePS.1...3.
M1.T2Rang
ePS.1...3.
M1.T2Rang
ePS.1...3.
M1.T2Rang
ePS.1...3.
L-2 1 -1,67197 -1,91224 -1,23188 -0,929507 -1,00966 -0,541067 -0,187641 -0,558269 -0,143506 -0,836282
L-3 2 -1,66196 -1,42348 -1,47412 -0,309718 -0,766982 -0,537983 -0,830685 -0,863842 -1,08623 -1,50687
L-5 3 -1,15128 -1,52646 -1,47608 -1,38188 -1,52171 -1,14752 -1,41737 -1,29394 -1,34483
L-6 4 0,0626525 -0,392714 -0,266382 -0,808636 -0,298793 -0,323792 -0,433484 -0,451272 -0,387277 -0,357159
L-7 5 0,992867 -0,0612148 -0,0845685 0,0785673 0,373838 0,357161 0,537244 0,51752 0,716233 0,740481
L-11 6 -0,13474 0,426421 -0,687178 -0,912154 -0,727428 -0,594267 -0,215358 -0,244603 -0,276336 -0,0408475
L-13 7 0,956142 1,29953 1,25385 1,44639 1,1509 1,05323 0,579315 0,791941 0,762827 0,452327
L-14 8 0,3823 0,979317 1,68164 2,11873 1,9835 2,03344 2,09829 1,71106 1,80616 1,71117
L-15 9 -1,11901 -1,04536 -0,910187 -0,16009 -0,384889 -0,556257 -0,48036 -0,579415 -0,47443 -0,972214
L-16 10 0,296386 0,551828 0,79582 0,750663 0,439351 0,248163 0,198917 0,145894 0,297474 0,312267
L-18 11 0,311971 0,531029 0,0926541 0,364508 0,339701 -0,440982 -0,522234 -0,349162 -0,672595 0,0737407
L-19 12 -0,976239 -0,364278 -0,0345314 0,0443366 -0,323212 -1,07351 -0,798123 -0,562394 -0,328668 -0,582896
L-21 13 0,812092 0,596723 1,46484 1,39453 1,5312 1,84158 1,79598 1,79942 1,60244 1,3785
L-23 14 1,17534 0,805715 0,0761837 -0,185552 0,364449 0,760569 0,932098 1,52143 1,37599 1,46404
L-108 15 1,11653 0,810736 0,0555947 -1,08214 0,249403 0,168306 0,0574737 -0,254304 -0,495755 -0,692051
L-110 16 0,608923 0,724441 0,74435 -0,428049 -1,39967 -1,24707 -1,31406 -1,33007 -1,35151 -1,14421
Risk_Index_
M1_14
M1
14
PAI
0,486223
0,731464
-0,87203
-0,87203
-0,360617
-0,431795
-0,87203
1,42551
-0,87203
-0,87203
0,832052
-0,87203
1,36975
1,99633
0,0552916
-0,87203